Method for treating cancer using chemokine antagonists

ABSTRACT

What is described is a method for treating cancer in a patient assaying levels of (a) CXCR1 and/or CXCR2 ligands or (b) myeloid derived suppressor cells (MDSCs) and/or neutrophils in the patient, and if the patient has increased levels of (a) or (b), then administering a therapeutically effective amount of a compound having a structure selected from the group consisting of formulas SX-517, SX-576, and SX-682

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/354,936 filed on Nov. 17, 2016, which issued as U.S. Pat. No.10,046,002, on Aug. 14, 2018. U.S. patent application Ser. No.15/279,361 filed on Sep. 28, 2016, U.S. patent application Ser. No.14/610,960 filed on Jan. 30, 2015, now issued U.S. Pat. No. 9,480,694,and U.S. patent application Ser. No. 13/957,665, filed Aug. 2, 2013, nowissued U.S. Pat. No. 8,969,365, are incorporated herein by reference.

GOVERNMENT RIGHTS

This invention was made with government support under Grant NumberHL072614 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

TECHNICAL FIELD

The present disclosure provides a method of treating cancer usingpyrimidinecarboxamide compounds, and pharmaceutical compositions thatinclude these compounds.

BACKGROUND

Chemokines are chemotactic proteins that have the potential to attractmacrophages, T-cells, eosinophils, basophils, neutrophils andendothelial cells to sites of inflammation and tumor growth. Chemokinesare typically low molecular mass (7-9 kD) proteins that can be dividedinto four subfamilies: CC (or β-chemokines), CXC, C (or γ-chemokines)and CX3C (or δ-chemokines). The chemokines are categorized through theirprimary amino acid structure. The CXC subfamily is characterized by twoconserved Cys residues (C) near the N-terminus and separated by an aminoacid (X). The CXC-chemokines include, for example, interleukin-8 (IL-8),neutrophil-activating protein-1 (NAP-1), neutrophil-activating protein-2(NAP-2), GROα, GROβ, GROγ, ENA-78, GCP-2, IP-10, MIG and PF4. The CXCsubfamily of chemokines is further characterized by the presence orabsence of a specific amino acid sequence, glutamicacid-leucine-arginine (or ELR for short) immediately before the firstCys residue of the CXC motif Those chemokines with the ELR motif(ELRCXC) are important for the recruitment and activation of neutrophilsto sites of inflammation. GROα and IL-8 are examples of ELRCXCchemokines.

The CXC-chemokines mediate their chemotactic activity throughinteraction with the chemokine receptors CXCR1 and CXCR2. CXCR1 bindsIL-8 and GCP-2 with high affinity while CXCR2 binds all ELRCXCchemokines with high affinity.

Since CXC-chemokines promote the accumulation and activation ofneutrophils, CXC-chemokines have been implicated in a wide range ofacute and chronic inflammatory disorders including COPD, psoriasis andrheumatoid arthritis. (Baggiolini, 1992, FEBS Lett, 307:97; Miller,1992. Crit Rev Immuno., 12:17; Oppenheim, 1991, Annu Rev Immunol, 9:617;Seitz, 1991, J Clin Invest, 87:463; Miller, 1992, Am Rev Respir Dis,146:427; Donnely, 1998, Lancet, 341:643).

ELRCXC chemokines, including IL-8, GROα, GROβ, GROγ, NAP-2, and ENA-78(Strieter, 1995, J Biol Chem, 270:27348-57), have also been implicatedin the induction of tumor angiogenesis (new blood vessel growth).Angiogenic activity is due to ELRCXC-chemokine binding to, andactivation of CXCR2, and possibly CXCR1 for IL-8, expressed on thesurface of vascular endothelial cells (ECs) in surrounding vessels.

Many different types of tumors have been shown to produce ELRCXCchemokines. Chemokine production has been correlated with a moreaggressive phenotype (Inoue, 2000, Clin Cancer Res, 6:2104-2119) andpoor prognosis (Yoneda, 1998, J Nat Cancer Inst, 90:447-54). Chemokinesare potent chemotactic factors and the ELRCXC chemokines, in particular,have been shown to induce EC chemotaxis. Thus, these chemokines arethought to induce chemotaxis of endothelial cells toward their site ofproduction in the tumor. This may be a critical step in the induction ofangiogenesis by the tumor. Inhibitors of CXCR2 or dual inhibitors ofCXCR2 and CXCR1 will inhibit the angiogenic activity of the ELRCXCchemokines and therefore block the growth of the tumor. This anti-tumoractivity has been demonstrated for antibodies to IL-8 (Arenberg, 1996, JClin Invest, 97:2792-802), ENA-78 (Arenberg, 1998, J Clin Invest,102:465-72), and GROα (Haghnegandar, 2000, J Leukoc Biology, 67:53-62).

Therefore, there is a need in the art to find CXCR1/2 inhibitorcompounds and modulator compounds that can be used as pharmaceuticalcompounds. There remains a need for compounds that are capable ofmodulating activity at CXC-chemokine receptors. For example, conditionsassociated with an increase in IL-8 production (which is responsible forchemotaxis of neutrophil and T-cell subsets into the inflammatory siteand growth of tumors) would benefit by compounds that are inhibitors ofIL-8 receptor binding. The present disclosure was made to satisfy thisneed.

SUMMARY

The present disclosure provides a compound having the formula SX-517,SX576, or SX-682.

The present disclosure further provides a pharmaceutical compositioncomprising a compound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt, or solvate thereof and apharmaceutically acceptable carrier. In certain embodiments, thisdisclosure provides SX-517, SX576, or SX-682 as novel compounds that areCXC chemokine-modulators, pharmaceutical compositions comprising acompound having the formula SX-517, SX576, or SX-682, and methods oftreatment, prevention, inhibition, or amelioration of one or morediseases associated with CXC chemokine mediation using a compound havingthe formula SX-517, SX576, or SX-682, and compositions disclosed herein.

The present disclosure provides a method for treating a disease ordisorder selected from the group consisting of pain (e.g., acute pain,acute inflammatory pain, chronic inflammatory pain, and neuropathicpain), acute inflammation, chronic inflammation, rheumatoid arthritis,psoriasis, atopic dermatitis, asthma, bronchopulmonary dysplasia, COPD,adult respiratory disease, arthritis, inflammatory bowel disease,Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, stroke, ischemia reperfusioninjury, renal reperfusion injury, glomerulonephritis, thrombosis,Alzheimer's disease, graft vs. host reaction (i.e., graft-versus-hostdisease), allograft rejections (e.g., acute allograft rejection, andchronic allograft rejection), malaria, acute respiratory distresssyndrome, delayed type hypersensitivity reaction, atherosclerosis,cerebral ischemia, cardiac ischemia, osteoarthritis, multiple sclerosis,restinosis, angiogenesis, angiogenesis associated with tumor growth,osteoporosis, gingivitis, respiratory viruses, herpes viruses, hepatitisviruses, HIV, Kaposi's sarcoma associated virus (i.e., Kaposi'ssarcoma), meningitis, cystic fibrosis, pre-term labor, cough, pruritis,multi-organ dysfunction, trauma, strains, sprains, contusions, psoriaticarthritis, herpes, encephalitis, CNS vasculitis, traumatic brain injury,systemic tumors, CNS tumors, tumors dependent on angiogenesis forgrowth, leukopenia and neutropenia, chemotherapy-induced leukopenia andneutropenia, opportunistic infections associated with neutropenia orleukopenia, subarachnoid hemorrhage, post-surgical trauma, interstitialpneumonitis, hypersensitivity, crystal induced arthritis, acutepancreatitis, chronic pancreatitis, acute alcoholic hepatitis,necrotizing enterocolitis, chronic sinusitis, angiogenic ocular disease,ocular inflammation, retinopathy of prematurity, diabetic retinopathy,macular degeneration with the wet type preferred, cornealneovascularization, polymyositis, vasculitis, acne, gastric ulcers,duodenal ulcers, celiac disease, esophagitis, glossitis, airflowobstruction, airway hyperresponsiveness (i.e., airway hyperreactivity),bronchiectasis, bronchiolitis, bronchiolitis obliterans, chronicbronchitis, cor pulmonae, dyspnea, emphysema, hypercapnea,hyperinflation, hypoxemia, hyperoxia-induced inflammations, hypoxia,surgical lung volume reduction, pulmonary fibrosis, pulmonaryhypertension, right ventricular hypertrophy, peritonitis associated withcontinuous ambulatory peritoneal dialysis (CAPD), granulocyticehrlichiosis, sarcoidosis, small airway disease, ventilation-perfusionmismatching, wheeze, colds, gout, alcoholic liver disease, lupus, burntherapy (i.e., the treatment of burns), periodontitis, cancer,transplant reperfusion injury, and early transplantation rejection(e.g., acute allograft rejection) in a patient in need of suchtreatment, comprising administering an effective amount of the compoundhaving the formula SX-682.

The present disclosure further provides a method for treating a patientwith a cancer, comprising assaying levels of (a) CXCR1 and/or CXCR2ligands or (b) myeloid derived suppressor cells (MDSCs) and/orneutrophils in a patient with a cancer, and if the patient has increasedlevels of (a) or (b), wherein the increased levels are caused by thecancer, then administering a therapeutically effective amount of acompound having a structure selected from the group consisting offormulas SX-517, SX-576, and SX-682. The method further comprisesadministering an anticancer therapy, e.g., wherein the anticancertherapy is a chemotherapy, including wherein carboplatin is administeredto the patient. The method further comprises wherein the anticancertherapy is radiation therapy.

In one embodiment the method uses the compound having the formulaSX-517, SX576, or SX-682 and the cancer is selected from cancers of theprostate, colorectum, pancreas, cervix, stomach, endometrium, brain,liver, bladder, ovary, testis, head, neck, skin, mesothelial lining,blood, esophagus, breast, muscle, connective tissue, lung, adrenalgland, thyroid, kidney, lymphoid system, bone marrow or bone,glioblastoma, mesothelioma, renal cell carcinoma, gastric carcinoma,sarcoma, choriocarcinoma, cutaneous basocellular carcinoma, andtesticular seminoma.

In another embodiment the method uses the compound having the formulaSX-517, SX576, or SX-682 and the cancer is selected from cancers of theprostate, pancreas, stomach, brain, liver, head, neck, skin, blood,breast, lung, glioblastoma, and mesothelioma.

In another embodiment the method uses the compound having the formulaSX-517, SX576, or SX-682 and the cancer is breast cancer, preferably byoral administration. The breast cancer therapy further comprisesadministering a platinum chemotherapy.

In another embodiment the method uses the compound having the formulaSX-517, SX576, or SX-682 and the cancer is a melanoma, preferably byoral administration. The melanoma cancer treatment further comprisesadministering a platinum chemotherapy.

In another embodiment, the method uses the compound of formula SX-517,SX-576, or SX-682, for treating a lung cancer, preferably by oraladministration.

In another embodiment, the method uses the compound has the compoundhaving the formula SX-517, SX576, or SX-682 for treating a prostatecancer, preferably by oral administration.

In another embodiment, the method uses the compound of formula SX-517,SX-576, or SX-682, for treating a cancer, wherein the cancer is aglioblastoma, preferably by oral administration. The glioblastomatreatment further comprises administering temozolomide.

In another embodiment, the method uses the compound of formula SX-517,SX-576, or SX-682, for treating pancreatic cancer, preferably by oraladministration.

The present disclosure further provides a method for treating a patientwith a cancer, comprising administering to the patient in need thereof apharmaceutical composition comprising a therapeutically effective amountof a compound having a structure selected from the group consisting offormulas SX-517, SX-576, and SX-682, or a pharmaceutically

acceptable salt or solvate thereof, wherein the cancer is a glioblastomaor a pancreatic cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows that SX-682 inhibited CXCL8-mediated intracellular calciumflux in isolated human neutrophils (‘Human PMNs’), RBL cells stablytransfected with CXCR1 (legend ‘CXCR1’), and RBL cells stablytransfected with CXCR2 (legend ‘CXCR2’). Mean (n=4, ±SE) IC₅₀ values forSX-682 in each cell system are in parentheses in the legend.

FIG. 2 shows that inhibition of CXCL8-mediated intracellular calciumflux in RBL cells stably transfected with CXCR1 is sustained for atleast 12 hours after SX-682 washout.

FIG. 3 shows that inhibition of CXCL8-mediated intracellular calciumflux in RBL cells stably transfected with CXCR2 is sustained for atleast 12 hours after SX-682 washout.

FIG. 4 shows the effect of intravenous dosing of either SX-576 or SX-682on neutrophil influx in the ozone rat model of pulmonary inflammation.

FIG. 5 illustrates boronic acid containing CXCR1/CXCR2 inhibitors.

FIG. 6 shows structures of selected antagonists of CXCR1 and/or CXCR2 offormula I that include SX-517, SX-520, SX-557, SX-574, SX-577, SX-603,SX-622, SX-660, SX-662, SX-671, SX-677, SX-678, and SX-682.

FIG. 7 shows the results of inhibition of myeloid derived suppressorcells (MDSCs) migration by SX-682 in a dose-dependent fashion. The graphshows SX-682 concentration, micromolar (μmolar), vs. the number ofmigrating MDSCs as measured by flow cytometry. As a control, an aqueoussolution with carrier solvent DMSA was used (0 μmolar).

FIG. 8A shows the results of direct inhibition of tumor cellproliferation of leukemia cell lines by SX-682 in a dose-dependentfashion. The leukemia cell lines included CCRF-CEM, MOLT-4, HL-60,RPMI-8226, K-562 and SR. These were independently culture in grownmedium at five concentrations of SX-682 between 10⁻⁸ M to 10⁻⁴ molar.Percentage growth inhibition was measured by dye adsorption.

FIG. 8B shows the results of direct inhibition of tumor cellproliferation of non-small cell lung cancer cell lines by SX-682 in adose-dependent fashion as measured in FIG. 7A. The non-small cell lungcancer cell lines included A549, H226, H460, H23, H522, H322M, HOP-62,and HOP-92.

FIG. 8C shows the results of direct inhibition of tumor cellproliferation of colon cancer cell lines by SX-682 in a dose-dependentfashion as measured in FIG. 7A. The colon cancer cell lines includedCOLO 205, HCT-15, HCC-2998, KM12, HCT-116, and SW-620.

FIG. 8D shows the results of direct inhibition of tumor cellproliferation of CNS cancer cell lines by SX-682 in a dose-dependentfashion as measured in FIG. 7A. The CNS cancer cell lines includedSF-268, SNB-19, SF-295, SNB-75, and SF-539.

FIG. 8E shows the results of direct inhibition of tumor cellproliferation melanoma cell lines by SX-682 in a dose-dependent fashion.The melanoma cell lines included LOX IMVI, MDA-MB-435, SK-MEL-5,MALME-3M, SK-MEL-2, UACC-257, M14, SK-MEL-28, and UACC-62.

FIG. 8F shows the results of direct inhibition of tumor cellproliferation of ovarian cancer cell lines by SX-682 in a dose-dependentfashion. The ovarian cancer cell lines included IBROV1, OVCAR-8,OVCAR-3, NCl/ADR-RES, OVCAR-5, and SK-OV-3.

FIG. 8G shows the results of direct inhibition of tumor cellproliferation of renal cancer cell lines by SX-682 in a dose-dependentfashion. The renal cancer cell lines included 786-0, CAK-1, TK-10, RXF393, UO-31, ACHN, and SN12C.

FIG. 8H shows the results of direct inhibition of tumor cellproliferation of prostate cancer cell lines by SX-682 in adose-dependent fashion. The prostate cancer cell lines included PC-3 andDU-145.

FIG. 8I shows the results of direct inhibition of tumor cellproliferation of breast cancer cell lines by SX-682 in a dose-dependentfashion. The breast cancer cell lines included MCF7, BT-549, MDA-MB-231,T-47D, HS 578T, and MDA-MB-468.

FIG. 9 shows that SX-682 alone or in combination with carboplatin (CP)was an effective therapy for breast cancer in a validated mouse model.The volume (mm³) of tumors in T11 genetically engineered mice wasmeasured during 1-2 week treatment by daily oral administration ofSX-682 (10 mg/day/mouse) with or without administration by weekly IPinjection of CP (50 mg/kg). The mean, SE, and statistical significanceof P<0.0001 (***) for cohorts of seven mice are shown.

FIG. 10 shows that SX-682 alone or synergistically in combination withCP was an effective treatment for breast cancer in a validated animalmodel. The volume (mm³) of tumors in C3Tag genetically engineered micewas measured during 4-6 week treatment dosing as in FIG. 9. The mean,SE, and statistical significance of P<0.0001 for cohorts of 10-12 miceare shown.

FIG. 11 shows that SX-682 synergized CP to improve survival in atreatment for breast cancer. The treatments were administered in theC3Tag genetically engineered mouse model of breast cancer as describedin FIG. 10. The treatment extended for up to 60 days. The results showthat the combination therapy (SX-682+CP) significantly improved survivalover CP or SX-682 alone (P=0.008).

FIG. 12 shows that SX-682 alone or in combination with CP was aneffective treatment for melanoma in a validated animal model. The volume(mm³) of tumors in TRIA genetically engineered mice was measured during15-20 week treatment dosing SX-682 and CP as in FIG. 9. The mean, SE,and statistical significance of P<0.0001 for cohorts of 10-12 mice areshown.

FIG. 13 shows that SX-682 synergized with carboplatin (CP) to causeremission of melanomas in the validated mouse model as described in FIG.12. 21-days of combination treatment caused a significant percentagechange in tumor volume in TRIA genetically engineered mice compared toCP treatment alone.

FIG. 14 shows that SX-682 alone or in combination with anti-PD-1antibody was effective in treating melanomas in a mouse model. The tumorarea (mm2) in B16-F10 syngeneic mice was measured during 26 days oftreatment with SX-682 orally administered twice daily (50 mg/kg) withand without 100 μg anti-PD-1 administered twice weekly by IP injection.SX-682 monotherapy significantly slowed tumor growth vs. control(P=0.0002, **) and synergized with anti-PD1 therapy vs. monotherapy witheither SX-786 or anti-PD-1 (P<0.0005, ***). Data and error bars are themean±SE of 4 or 5 mice per cohort.

FIG. 15 shows that SX-682 alone or in combination with anti-PD-1antibody inhibited lung cancer in a validated animal model using thedosing amounts as in FIG. 14. Tumor area (mm2) was measured in LLCsyngeneic mice. SX-682 monotherapy significantly slowed tumor growth vs.control (P=0.0002, **) and synergized with anti-PD1 therapy vs.monotherapy with either SX-682 or anti-PD-1 (P<0.002, ***). Data anderror bars are the mean±SE in cohorts of 5 mice.

FIG. 16 shows that SX-682 synergized with anti-PD-1 antibody to increasesurvival in the LLC syngeneic mice as described in FIG. 15. Drugadministration and survival determinations continued up to 66 days aftertumor injection. The combination of SX-682 and anti-PD-1 therapysignificantly enhanced survival compared to vehicle or anti-PD-1 therapyalone (P=0.002).

FIG. 17 shows that SX-682 alone and in combination with immunecheckpoint blockade (anti-PD1 and anti-CTLA4) inhibited prostate cancerin a validated animal model. Ptenpc−/−p53pc−/−Smad4pc−/− mice wereadministered 50 mg/kg SX-682 by oral gavage b.i.d. and 200 μg each ofanti-PD1 and anti-CTLA4 antibodies (ICB), 3×/week. Prostate weight (g)was measured after 4-6 weeks. SX-682 plus ICB was significantly betterthan control (P=0.0016, **), and ICB or SX682 alone (P=0.021, *)(unpaired t-test). Mean+SE are shown.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

When any substituent or variable occurs more than one time in anymoiety, its definition on each occurrence is independent of itsdefinition at every other occurrence. Also, combinations of substituentsand/or variables are permissible only if such combinations result instable compounds.

Unless indicated otherwise, the following definitions apply throughoutthe present specification and claims. These definitions apply regardlessof whether a term is used by itself or in combination with other terms.For example, the definition of “alkyl” also applies to the “alkyl”portion of the defined term “alkoxy”.

“An effective amount” or a “therapeutically effective amount” means todescribe an amount of compound of the present disclosure or anotheragent effective to treat a mammal (e.g., a human) having a disease orCXC chemokine-mediated condition, and thus producing the desiredtherapeutic effect.

“At least one” means one or more (e.g., 1-3, 1-2, or 1).

“Composition” includes a product comprising the specified ingredients inthe specified amounts, as well as any product that results, directly orindirectly, from combination of the specified ingredients in thespecified amounts.

“In combination with” as used to describe the administration of thecompound having the formula SX-517, SX576, or SX-682 with othermedicaments in the methods of treatment of this invention, means thatthe compound having the formula SX-517, SX576, or SX-682 and the othermedicaments are administered sequentially or concurrently in separatedosage forms, or are administered concurrently in the same dosage form.

“Mammal” means a human or other mammal or means a human being.

“Patient” includes both human and other mammals, preferably human.

“Prodrug” denotes a compound that is a drug precursor which, uponadministration to a subject, undergoes chemical conversion by metabolicor chemical processes to yield the compound having the formula SX-517,SX576, or SX-682 or a salt and/or solvate thereof. A discussion ofpro-drugs is provided in Higuchi, Pro-drugs as Novel Delivery Systems,v. 14 of the A.C.S. Symp Series, and in Bioreversible Carriers in DrugDesign, Roche, ed., Am Pharma Assoc, Pergamon Press, 1987, both of whichare incorporated herein by reference.

“Chemokine” means a protein molecule involved in chemotaxis.

A “chemokine-mediated disease” means a disease of which at least oneelement or cause is related to the regulation of a CXC chemokine.

“Commercially available chemicals” and the chemicals used in theExamples set forth herein may be obtained from standard commercialsources, where such sources include, for example, Acros Organics(Pittsburgh, Pa.), Sigma-Adrich Chemical (Milwaukee, Wis.), AvocadoResearch (Lancashire, U.K.), Bionet (Cornwall, U.K.), Boron Molecular(Research Triangle Park, N.C.), Combi-Blocks (San Diego, Calif.),Eastman Organic Chemicals, Eastman Kodak Company (Rochester, N.Y.),Fisher Scientific Co. (Pittsburgh, Pa.), Frontier Scientific (Logan,Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.), Lancaster Synthesis(Windham, N.H.), Maybridge Chemical Co. (Cornwall, U.K.), PierceChemical Co. (Rockford, Ill.), Riedel de Haen (Hannover, Germany),Spectrum Quality Product, Inc. (New Brunswick, N.J.), TCI America(Portland, Oreg.), and Wako Chemicals USA, Inc. (Richmond, Va.).

“Compounds described in the chemical literature” may be identifiedthrough reference books and databases directed to chemical compounds andchemical reactions, as known to one of ordinary skill in the art.Suitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds disclosed herein, orprovide references to articles that describe the preparation ofcompounds disclosed herein, include for example, “Synthetic OrganicChemistry”, John Wiley and Sons, Inc. New York; S. R. Sandler et al,“Organic Functional Group Preparations,” 2^(nd) Ed., Academic Press, NewYork, 1983; H. O. House, “Modern Synthetic Reactions,” 2^(nd) Ed., W. A.Benjamin, Inc. Menlo Park, Calif., 1972; T. L. Glichrist, “HeterocyclicChemistry,” 2^(nd) Ed. John Wiley and Sons, New York, 1992; J. March,“Advanced Organic Chemistry: reactions, Mechanisms and Structure,”5^(th) Ed., Wiley Interscience, New York, 2001; Specific and analogousreactants may also be identified through the indices of known chemicalsprepared by the Chemical Abstract Service of the American ChemicalSociety, which are available in most public and university libraries, aswell as through online databases (the American Chemical Society,Washington, D.C.). Chemicals that are known but not commerciallyavailable in catalogs may be prepared by custom chemical synthesishouses, where many of the standard chemical supply houses (e.g. thoselisted above) provide custom synthesis services.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the presentdisclosure effective in decreasing or increasing (i.e., modulating) theaction of a CXC chemokine at a CXC chemokine receptor and thus producingthe desired therapeutic effect in a suitable patient.

“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Solvate” means a physical association of a compound of this disclosurewith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances, the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.

Examples of “disease modifying antirheumatic drugs” (i.e., DMARDs)include, for example, methotrexate, aminopterin, sulfasalzine,leflunomide, TNFα directed agents (e.g., infliximab, etanercept, andadalimumab), IL-1 directed agents (e.g., anakinra) B cell directedagents (e.g., rituximab), T cell directed agents (e.g., alefacept,efalizumab, and CTLA4-1g), TNFα-converting enzyme inhibitors,interleukin-1 converting enzyme is inhibitors, and p38 kinaseinhibitors.

The term “other classes of compounds indicated for the treatment ofrheumatoid arthritis”, as used herein, unless indicated otherwise,means: compounds selected from the group consisting of: IL-1 directedagents (e.g., anakinra); B cell directed agents (e.g., rituximab); Tcell directed agents (e.g., alefacept, efalizumab, and CTLA4-1g),TNFα-converting enzyme inhibitors, interleukin-1 converting enzymeinhibitors, and p38 kinase inhibitors.

A compound having the formula SX-517, SX576, or SX-682 forms salts thatare also within the scope of this disclosure. Reference to the compoundhaving the formula SX-517, SX576, or SX-682 herein is understood toinclude reference to salts thereof, unless otherwise indicated. The term“salt(s)”, as employed herein, denotes acidic salts formed withinorganic and/or organic acids, as well as basic salts formed withinorganic and/or organic bases. The salts can be pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts, althoughother salts are also useful. Salts of the compound having the formulaSX-517, SX576, or SX-682 may be formed, for example, by reacting it withan amount of acid or base, such as an equivalent amount, in a mediumsuch as one in which the salt precipitates or in an aqueous mediumfollowed by lyophilization.

Exemplary acid addition salts include acetates, adipates, alginates,ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates,borates, butyrates, citrates, camphorates, camphorsulfonates,cyclopentanepropionates, digluconates, dodecylsulfates,ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates,hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides,hydroiodides, 2-hydroxy ethanesulfonates, lactates, maleates,methanesulfonates, 2-napthalenesulfonates, nicotinates, nitrates,oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates,picrates, pivalates, propionates, salicylates, succinates, sulfates,sulfonates (such as those mentioned herein), tartarates, thiocyanates,toluenesulfonates (also known as tosylates) undecanoates, and the like.Additionally, acids which are generally considered suitable for theformation of pharmaceutically useful salts from basic pharmaceuticalcompounds are discussed, for example, by Berge, 1977, J. Pharma Sci,66:1-19; Gould, 1986, Int'l Pharmaceutics, 33:201-17; Anderson, ThePractice of Medicinal Chemistry (1996), Acad Press, New York; and in TheOrange Book (Food & Drug Administration, Washington, D.C.). Thesedisclosures are incorporated herein by reference herein.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salts with organic bases (for example,organic amines) such as benzathines, dicyclohexylamines, hydrabamines(formed with N,N-bis(dehydroabietyl)ethylenediamine),N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, and saltswith amino acids such as arginine, lysine and the like. Basicnitrogen-containing groups may be quarternized with agents such as loweralkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides,bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl,dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl,myristyl, and stearyl chlorides, bromides and iodides), arylalkylhalides (e.g., benzyl and phenethyl bromides), and others.

All such acid and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the disclosure and all acid andbase salts are considered equivalent to the free forms of thecorresponding compounds for purposes of the disclosure.

The compound having the formula SX-517, SX576, or SX-682 can exist inunsolvated and solvated forms, including hydrated forms. In general, thesolvated forms, with pharmaceutically acceptable solvents such as water,ethanol and the like, are equivalent to the unsolvated forms for thepurposes of this disclosure.

The compound having the formula SX-517, SX576, or SX-682 and salts,solvates and prodrugs thereof, may exist in their tautomeric form (forexample, as an amide or imino ether). All such tautomeric forms arecontemplated herein as part of the present disclosure.

Also, within the scope of the present disclosure are polymorphs of thecompounds of this disclosure (i.e., polymorphs of the compound havingthe formula SX-517, SX576, or SX-682 are within the scope of thisdisclosure).

Prodrugs of the compound having the formula SX-517, SX576, or SX-682 orpharmaceutically acceptable salts or solvates thereof are within thescope of this disclosure.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the compound having the formula SX-517, SX576, or SX-682(including those of the salts, solvates and prodrugs of the compounds aswell as the salts and solvates of the prodrugs), such as those which mayexist due to asymmetric carbons on various substituents, includingenantiomeric forms (which may exist even in the absence of asymmetriccarbons), rotameric forms, atropisomers, and diastereomeric forms, arecontemplated within the scope of this disclosure. Individualstereoisomers of the compounds of this disclosure may, for example, besubstantially free of other isomers, or may be admixed, for example, asracemates or with all other, or other selected, stereoisomers. Thechiral centers of the compounds herein can have the S or R configurationas defined by the IUPAC 1974 Recommendations. The use of the terms“salt”, “solvate”, “prodrug” and the like, is intended to equally applyto the salt, solvate and prodrug of enantiomers, stereoisomers,rotamers, tautomers, racemates or prodrugs of the disclosed compounds.

Classes of compounds that can be used as the chemotherapeutic agent(antineoplastic agent) include: alkylating agents, antimetabolites,natural products and their derivatives, hormones and steroids (includingsynthetic analogs), and synthetics. Examples of compounds within theseclasses are given below.

Alkylating agents (including nitrogen mustards, ethyleniminederivatives, alkyl sulfonates, nitrosoureas and triazenes): uracilmustard, chlormethine, cyclophosphamide, ifosfamide, melphalan,chlorambucil, pipobroman, triethylene-melamine,triethylenethiophos-phoramine, busulfan, carmustine, lomustine,streptozocin, dacarbazine, and temozolomide.

Antimetabolites (including folic acid antagonists, pyrimidine analogs,purine analogs and adenosine deaminase inhibitors): methotrexate,aminopterin, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine,6-thioguanine, fludarabine phosphate, pentostatine, and gemcitabine.

Natural products and their derivatives (including vinca alkaloids,antitumor antibiotics, enzymes, lymphokines and epipodophyllotoxins):vinblastine, vincristine, vindesine, bleomycin, dactinomycin,daunorubicin, doxorubicin, epirubicin, idarubicin, paclitaxel (TAXOL®),mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, interferons(especially IFN-γ), etoposide, and teniposide.

Hormones and steroids (including synthetic analogs):17α-ethinylestradiol, diethylstilbestrol, testosterone, prednisone,fluoxymesterone, dromostanolone propionate, testolactone,megestrolacetate, tamoxifen, methylprednisolone, methyltestosterone,prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone,aminoglutethimide, estramustine, medroxyprogesteroneacetate, leuprolide,flutamide, toremifene, zoladex.

Synthetics (including inorganic complexes such as platinum coordinationcomplexes): cisplatin, carboplatin, hydroxyurea, amsacrine,procarbazine, mitotane, mitoxantrone, levamisole, andhexamethylmelamine.

Methods for the safe and effective administration of most of thesechemotherapeutic agents are known to those skilled in the art. Inaddition, their administration is described in the standard literature.For example, the administration of many of the chemotherapeutic agentsis described in the “Physicians' Desk Reference” (PDR), e.g., 2008edition (Thomson P D R, Montvale, N.J. 07645); the disclosure of whichis incorporated herein by reference herein.

As used herein, a microtubule affecting agent is a compound thatinterferes with cellular mitosis, i.e., having an anti-mitotic effect,by affecting microtubule formation and/or action. Such agents can be,for instance, microtubule stabilizing agents or agents that disruptmicrotubule formation.

Microtubule affecting agents useful in this disclosure are well known tothose of skilled in the art and include, but are not limited toallocolchicine (NSC 406042), halichondrin B (NSC 609395), colchicine(NSC 757), colchicine derivatives (e.g., NSC 33410), dolastatin 10 (NSC376128), maytansine (NSC 153858), rhizoxin (NSC 332598), paclitaxel(TAXOL®, NSC 125973), TAXOL® derivatives (e.g., derivatives (e.g., NSC608832), thiocolchicine (NSC 361792), trityl cysteine (NSC 83265),vinblastine sulfate (NSC 49842), vincristine sulfate (NSC 67574),epothilone A, epothilone, and discodermolide (see Service, 1996,Science, 274:2009) estramustine, nocodazole, MAP4, and the like.Examples of such agents are also described in the scientific and patentliterature, see, e.g., Bulinski, 1997, J Cell Sci, 110:3055-64; Panda,1997, Proc Natl Acad Sci USA, 94:10560-64; Muhlradt, 1997, Cancer Res,57:3344-46; Nicolaou, 1997, Nature, 387:268-72; Vasquez, 1997, Mol BiolCell, 8:973-85; and Panda, 1996, J Biol Chem, 271:29807-12.

Particularly, agents can be compounds with paclitaxel-like activity.These include but are not limited to paclitaxel and paclitaxelderivatives (paclitaxel-like compounds) and analogues. Paclitaxel andits derivatives are available commercially. In addition, methods ofmaking paclitaxel and paclitaxel derivatives and analogues are wellknown to those of skilled in the art (see, e.g., U.S. Pat. Nos.5,569,729; 5,565,478; 5,530,020; 5,527,924; 5,508,447; 5,489,589;5,488,116; 5,484,809; 5,478,854; 5,478,736; 5,475,120; 5,468,769;5,461,169; 5,440,057; 5,422,364; 5,411,984; 5,405,972; and 5,296,506,the disclosures of which are incorporated by reference herein).

Additional microtubule affecting agents can be assessed using one ofmany such assays known in the art, e.g., a semiautomated assay whichmeasures the tubulin-polymerizing activity of paclitaxel analogs incombination with a cellular assay to measure the potential of thesecompounds to block cells in mitosis (see Lopes, 1997, Cancer ChemotherPharmacol, 41:37-47).

Therapeutic Activity

Modulators of neutrophil activity can have great therapeutic benefit ina number of indications. In disease states characterized by animproperly heightened neutrophil response, an inhibitor of neutrophilactivity would be indicated. In patients suffering from, for exampleneutropenia, a neutrophil agonist or activator has clinical benefit. Invivo evaluation of two lead compounds SX-517 and SX-576 in the murineair-pouch model of inflammation, revealed that both inhibitory andagonist activity on neutrophils were achieved, depending on the dosegiven.

Methods of Treatment

One embodiment is directed to a pharmaceutical composition comprisingSX-682 or a pharmaceutically acceptable salt or solvate thereof, incombination with a pharmaceutically acceptable carrier.

The methods of treatment of this disclosure are advantageous in treatingdiseases where the ELR-CXC chemokine binds to CXCR2. Another embodimentof the disclosure is directed to a method of treating CXCR1/2 chemokinemediated diseases in a patient in need of such treatment comprisingadministering to the patient a therapeutically effective amount of thecompound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt or solvate thereof.

Another embodiment of the disclosure is a method of treating CXCR1/2chemokine mediated diseases in a patient in need thereof comprisingadministering to the patient (a) an effective amount of the compoundhaving the formula SX-517, SX576, or SX-682, or a pharmaceuticallyacceptable salt or solvate thereof, concurrently or sequentially with(b) at least one additional agent, drug, medicament, antibody and/orinhibitor useful for the treatment of CXCR1/2 chemokine mediateddiseases. Examples of the additional medicament, drug or agent include,but are not limited to, disease modifying antirheumatic drugs;nonsteroidal antiinflammatory drugs (NSAIDs); COX-2 selectiveinhibitors; COX-1 inhibitors; immunosuppressives; steroids; biologicalresponse modifiers; and other anti-inflammatory agents or therapeuticsuseful for the treatment of CXCR1/2 chemokine mediated diseases.

Another embodiment of the method of treating a CXCR1/2 chemokinemediated disease is administering (a) a therapeutically effective amountof the compound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt or solvate thereof, concurrently orsequentially with (b) at least one medicament selected from the groupconsisting of: disease modifying antirheumatic drugs; nonsteroidalanti-inflammatory drugs; COX-2 selective inhibitors; COX-1 inhibitors;immunosuppressives; steroids; biological response modifiers; and otheranti-inflammatory agents or therapeutics useful for the treatment ofCXCR1 and/or CXCR2 chemokine mediated diseases.

Another embodiment of this disclosure is a method for treating cancer ina patient in need of such treatment, the method comprises administeringto said patient a therapeutically effective amount of the compoundhaving the formula SX-517, SX576, or SX-682, or a pharmaceuticallyacceptable salt or solvate thereof. Another embodiment of thisdisclosure is a method for treating cancer comprising administering tothe patient a therapeutic amount of the compound having the formulaSX-517, SX576, or SX-682, or a pharmaceutically acceptable salt orsolvate thereof, concurrently or sequentially with (a) at least oneantineoplastic agent selected from the group consisting of: (1)gemcitabine, (2) paclitaxel, (3) 5-fluorouracil (5-FU), (4)cyclo-phosphamide, (5) temozolomide and (6) vincristine or (b) at leastone agent selected from the group consisting of (1) microtubuleaffecting agents, (2) antineoplastic agents, (3) anti-angiogenesisagents, (4) VEGF receptor kinase inhibitors, (5) antibodies against theVEGF receptor, (6) interferon, and (7) radiation.

This disclosure also provides a method for treating CXCR1/2 mediateddisease or condition selected from the group consisting of: pain (e.g.,acute pain, acute inflammatory pain, chronic inflammatory pain, andneuropathic pain), acute inflammation, chronic inflammation, rheumatoidarthritis, psoriasis, atopic dermatitis, asthma, bronchopulmonarydysplasia, COPD, adult respiratory disease, arthritis, inflammatorybowel disease, Crohn's disease, ulcerative colitis, septic shock,endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke,ischemia reperfusion injury, renal reperfusion injury,glomerulonephritis, thrombosis, Alzheimer's disease, graft vs. hostreaction (i.e., graft-versus-host disease), allograft rejections (e.g.,acute allograft rejection, and chronic allograft rejection), malaria,acute respiratory distress syndrome, delayed type hypersensitivityreaction, atherosclerosis, cerebral ischemia, cardiac ischemia,osteoarthritis, multiple sclerosis, restinosis, angiogenesis,angiogenesis associated with tumor growth, osteoporosis, gingivitis,respiratory viruses, herpes viruses, hepatitis viruses, HIV, Kaposi'ssarcoma associated virus (i.e., Kaposi's sarcoma), meningitis, cysticfibrosis, pre-term labor, cough, pruritis, multi-organ dysfunction,trauma, strains, sprains, contusions, psoriatic arthritis, herpes,encephalitis, CNS vasculitis, traumatic brain injury, systemic tumors,CNS tumors, tumors dependent on angiogenesis for growth, leukopenia andneutropenia, chemotherapy-induced leukopenia and neutropenia,opportunistic infections associated with neutropenia or leukopenia,subarachnoid hemorrhage, post-surgical trauma, interstitial pneumonitis,hypersensitivity, crystal induced arthritis, acute pancreatitis, chronicpancreatitis, acute alcoholic hepatitis, necrotizing enterocolitis,chronic sinusitis, angiogenic ocular disease, ocular inflammation,retinopathy of prematurity, diabetic retinopathy, macular degenerationwith the wet type preferred, corneal neovascularization, polymyositis,vasculitis, acne, gastric ulcers, duodenal ulcers, celiac disease,esophagitis, glossitis, airflow obstruction, airway hyperresponsiveness(i.e., airway hyperreactivity), bronchiectasis, bronchiolitis,bronchiolitis obliterans, chronic bronchitis, cor pulmonae, dyspnea,emphysema, hypercapnea, hyperinflation, hypoxemia, hyperoxia-inducedinflammations, hypoxia, surgical lung volume reduction, pulmonaryfibrosis, pulmonary hypertension, right ventricular hypertrophy,peritonitis associated with continuous ambulatory peritoneal dialysis(CAPD), granulocytic ehrlichiosis, sarcoidosis, small airway disease,ventilation-perfusion mismatching, wheeze, colds, gout, alcoholic liverdisease, lupus, burn therapy (i.e., the treatment of burns),periodontitis, cancer, transplant reperfusion injury, earlytransplantation rejection (e.g., acute allograft rejection) in a patientin need of such treatment comprising administering to said patient aneffective amount of the compound having the formula SX-517, SX576, orSX-682, or a pharmaceutically acceptable salt or solvate thereof.

Another embodiment of this disclosure is a method for treating diseasessuch as allograft rejections, early transplantation rejections,autoimmune deafness, myocarditis, neuropathies, autoimmune diseases andvasculitis syndromes wherein said:

-   -   (a) allograft rejections are selected from the group consisting        of acute allograft rejections and chronic allograft rejections;    -   (b) early transplantation rejection is an acute allograft        rejection; (c) autoimmune deafness is Meniere's disease;    -   (d) myocarditis is viral myocarditis;    -   (e) neuropathies are selected from the group consisting of IgA        neuropathy, membranous neuropathy and idiopathic neuropathy;    -   (f) autoimmune diseases are anemias; and (g) vasculitis        syndromes are selected from the group consisting of giant cell        arteries, Behcet's disease and Wegener's granulomatosis.

Another embodiment of this disclosure is a method for treating COPD in apatient in need of such treatment comprising administering to saidpatient a therapeutically effective amount of the compound having theformula SX-517, SX576, or SX-682, or a pharmaceutically acceptable saltor solvate thereof.

Another embodiment of this disclosure is a method for treating arthritisin a patient in need of such treatment comprising administering to saidpatient a therapeutically effective amount of the compound having theformula SX-517, SX576, or SX-682, or a pharmaceutically acceptable saltor solvate thereof.

Another embodiment of this disclosure is a method for treatingosteoarthritis in a patient in need of such treatment comprisingadministering to said patient a therapeutically effective amount of thecompound having the formula SX-517, SX576, or SX-682 2, or apharmaceutically acceptable salt or solvate thereof.

Another embodiment of this disclosure is a method for treating pain in apatient in need of such treatment comprising administering to saidpatient a therapeutically effective amount of the compound having theformula SX-517, SX576, or SX-682, or a pharmaceutically acceptable saltor solvate thereof.

Another embodiment of this disclosure is a method for treating pain in apatient in need of such treatment comprising administering to saidpatient a therapeutically effective amount of the compound having theformula SX-517, SX576, or SX-682, or a pharmaceutically acceptable saltor solvate thereof, and administering a therapeutically effective amountof at least one medicament selected from the group consisting of:NSAIDs, COXIB inhibitors (e.g., COX-1 and COX-2 inhibitors),anti-depressants, and anti-convulsants.

Another embodiment of this disclosure is a method for treating acutepain in a patient in need of such treatment comprising administering tosaid patient a therapeutically effective amount of the compound havingthe formula SX-517, SX576, or SX-682, or a pharmaceutically acceptablesalt or solvate thereof.

Another embodiment of this disclosure is a method for treating acuteinflammatory pain in a patient in need of such treatment comprisingadministering to said patient a therapeutically effective amount of thecompound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt or solvate thereof.

Another embodiment of this disclosure is a method for treating chronicinflammatory pain in a patient in need of such treatment comprisingadministering to said-patient a therapeutically effective amount of thecompound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt or solvate thereof.

Another embodiment of this disclosure is a method for treatingneuropathic pain in a patient in need of such treatment comprisingadministering to said patient a therapeutically effective amount of thecompound having the formula SX-517, SX576, or SX-682, or apharmaceutically acceptable salt or solvate thereof.

Another embodiment of this disclosure is a pharmaceutical compositioncomprising the compound having the formula SX-517, SX576, or SX-682, ora pharmaceutically acceptable salt or solvate thereof, and at least oneother agent, medicament, antibody and/or inhibitor disclosed above, anda pharmaceutically acceptable carrier.

In general, the compounds used to treat pain will have CXCR1/2antagonistic activity.

NSAIDs are well known to those skilled in the art and can be used intheir known dosages and dosage regimens. Examples of NSAIDs include butare not limited to: piroxicam, ketoprofen, naproxen, indomethacin, andibuprofen COXIB inhibitors are well known to those skilled in the artand can be used in their known dosages and dosage regimens. Examples ofCOXIB inhibitors include but are not limited to: rofecoxib andcelecoxib. Anti-depressants are well known to those skilled in the artand can be used in their known dosages and dosage regimens. Examples ofanti-depressants include but are not limited to: amitriptyline andnortriptyline. Anti-convulsants are well known to those skilled in theart and can be used in their known dosages and dosage regimens. Examplesof anti-convulsants include but are not limited to: gabapentin,carbamazepine, pregabalin, and lamotragine.

Pharmaceutical Compositions

For preparing pharmaceutical compositions from the compound having theformula SX-517, SX576, or SX-682, inert, pharmaceutically acceptablecarriers can be either solid or liquid. Solid form preparations includepowders, tablets, dispersible granules, capsules, cachets andsuppositories. The powders and tablets may be comprised of from about 5to about 95 percent active ingredient. Suitable solid carriers are knownin the art, e.g., magnesium carbonate, magnesium stearate, talc,microcrystalline cellulose, sugar or lactose. Tablets, powders, cachetsand capsules can be used as solid dosage forms suitable for oraladministration. Examples of pharmaceutically acceptable carriers andmethods of manufacture for various compositions may be found in A.Gennaro (ed.), Remington: The Science and Practice of Pharmacy, 20thEdition, (2000), Lippincott Williams & Wilkins, Baltimore, Md. which isincorporated herein by reference.

Liquid form preparations include solutions, suspensions and emulsions.For example, water or water-propylene glycol solutions are used forparenteral injection, or addition of sweeteners and opacifiers are usedfor oral solutions, suspensions and emulsions. Liquid form preparationsmay also include solutions for intranasal administration. Liquid formpreparations may also include dissolution in lipid-based,self-emulsifying drug delivery systems (SEDDS) such as LABRASOL® orGELUCIRE® for oral administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier, such as an inert compressed gas,e.g. nitrogen.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compound having the formula SX-517, SX576, or SX-682 may also bedeliverable transdermally. The transdermal composition can take the formof creams, lotions, aerosols and/or emulsions and can be included in atransdermal patch of the matrix or reservoir type as are conventional inthe art for this purpose.

The compound of formula SX-682 can be administered orally.

A suitable pharmaceutical preparation is in a unit dosage form. In suchform, the preparation is subdivided into suitably sized unit dosescontaining appropriate quantities of the active component, e.g., aneffective amount to achieve the desired purpose.

The quantity of the compound having the formula SX-517, SX576, or SX-682in a unit dose of preparation may be varied or adjusted from about 0.01mg to about 1000 mg, or from about 0.01 mg to about 750 mg, or fromabout 0.01 mg to about 500 mg, or from about 0.01 mg to about 250 mg,according to the particular application.

The actual dosage employed may be varied depending upon the requirementsof the patient and the severity of the condition being treated.Determination of the proper dosage regimen for a particular situation iswithin the skill of the art. For convenience, the total dosage may bedivided and administered in portions during the day as required.

The amount and frequency of administration of the compound having theformula SX-517, SX576, or SX-682 and/or the pharmaceutically acceptablesalts thereof will be regulated according to the judgment of theattending clinician considering such factors as age, condition and sizeof the patient as well as severity of the symptoms being treated. Atypical recommended daily dosage regimen for oral administration canrange from about 0.04 mg/day to about 4000 mg/day, in two to fourdivided doses, or given preferably as a single once-daily dose.Once-weekly and twice-weekly dosing is also preferable.

The amount and frequency of administration of the compound having theformula SX-517, SX576, or SX-682 and the chemotherapeutic agents and/orradiation therapy will be regulated according to the judgment of theattending clinician (physician) considering such factors as age,condition and size of the patient as well as severity of the diseasebeing treated. A dosage regimen of the compound having the formulaSX-517, SX576, or SX-682 can be orally administration of from 10 mg to2000 mg/day, or 10 to 1000 mg/day, or 50 to 600 mg/day, in two to four(or two) divided doses, to block tumor growth. Intermittent therapy(e.g., one week out of three weeks or three out of four weeks) may alsobe used.

The chemotherapeutic agent and/or radiation therapy can be administeredaccording to therapeutic protocols well known in the art. It will beapparent to those skilled in the art that the administration of thechemotherapeutic agent and/or radiation therapy can be varied dependingon the disease being treated and the known effects of thechemotherapeutic agent and/or radiation therapy on that disease. Also,in accordance with the knowledge of the skilled clinician, thetherapeutic protocols (e.g., dosage amounts and times of administration)can be varied in view of the observed effects of the administeredtherapeutic agents (i.e., antineoplastic agent or radiation) on thepatient, and in view of the observed responses of the disease to theadministered therapeutic agents.

If the compound having the formula SX-517, SX576, or SX-682, and thechemotherapeutic agent and/or radiation is not administeredsimultaneously or essentially simultaneously, then the initial order ofadministration of the compound having the formula SX-517, SX576, orSX-682, and the chemotherapeutic agent and/or radiation, may not beimportant. Thus, the compound having the formula SX-517, SX576, orSX-682 may be administered first, followed by the administration of thechemotherapeutic agent and/or radiation; or the chemotherapeutic agentand/or radiation may be administered first, followed by theadministration of the compound having the formula SX-517, SX576, orSX-682. This alternate administration may be repeated during a singletreatment protocol. The determination of the order of administration,and the number of repetitions of administration of each therapeuticagent during a treatment protocol, is well within the knowledge of theskilled physician after evaluation of the disease being treated and thecondition of the patient.

For example, the chemotherapeutic agent and/or radiation may beadministered first, especially if it is a cytotoxic agent, and then thetreatment continued with the administration of the compound having theformula SX-517, SX576, or SX-682 followed, where determinedadvantageous, by the administration of the chemotherapeutic agent and/orradiation, and so on until the treatment protocol is complete.

The particular choice of the compound having the formula SX-517, SX576,or SX-682, and chemotherapeutic agent and/or radiation will depend uponthe diagnosis of the attending physicians and their judgment of thecondition of the patient and the appropriate treatment protocol.

Also, in general, the compound having the formula SX-517, SX576, orSX-682 and the chemotherapeutic agent do not have to be administered inthe same pharmaceutical composition, and may, because of differentphysical and chemical characteristics, have to be administered bydifferent routes. For example, the compound of formula SX-682 may beadministered orally to generate and maintain good blood levels thereof,while the chemotherapeutic agent may be administered intravenously. Thedetermination of the mode of administration and the advisability ofadministration, where possible, in the same pharmaceutical composition,is well within the knowledge of the skilled clinician. The initialadministration can be made according to established protocols known inthe art, and then, based upon the observed effects, the dosage, modes ofadministration and times of administration can be modified by theskilled clinician.

Thus, in accordance with experience and knowledge, the practicingphysician can modify each protocol for the administration of a component(therapeutic agent; i.e., the compound having the formula SX-517, SX576,or SX-682, chemotherapeutic agent or radiation) of the treatmentaccording to the individual patient's needs, as the treatment proceeds.

The attending clinician, in judging whether treatment is effective atthe dosage administered, will consider the general well-being of thepatient as well as more definite signs such as relief of disease-relatedsymptoms, inhibition of tumor growth, actual shrinkage of the tumor, orinhibition of metastasis. Size of the tumor can be measured by standardmethods such as radiological studies, e.g., CAT or MRI scan, andsuccessive measurements can be used to judge whether or not growth ofthe tumor has been retarded or even reversed. Relief of disease-relatedsymptoms such as pain, and improvement in overall condition can also beused to help judge effectiveness of treatment.

The disclosure provided herein is exemplified by the followingpreparations and examples that should not be construed to limit thescope of the disclosure. For example, alternative mechanistic pathwaysand analogous structures may be apparent to those skilled in the art.

Pharmacology Example 1: In Vitro Inhibition of Intracellular CalciumRelease by SX-682

An in vitro assay showed inhibition of CXCR1/2-mediated intracellularcalcium release by SX-682. Briefly, cells (either isolated humanneutrophils or RBL cells stably transfected with either CXCR1 or CXCR2)were suspended in HBSS⁻ (without Ca²⁺ and Mg²⁺) containing 10 mM HEPESand FLIPR calcium 3 dye (3.1×10⁷ cells in total volume 1.7 mL). Cellswere aliquoted (200 μL of the cell suspension per tube, 8 tubes total)and 2 μL of the designated compound (with appropriate dilutions) wereadded to each of 6 tubes. As controls, 2 μL of DMSO (1% finalconcentration) were added to 2 other tubes. Cells were incubated for 30min at 37° C. After dye loading, tubes were centrifuged at 6,000 rpm for1 min, supernatant was removed, and the cell pellet was re-suspended in200 μL of HBSS⁺ (with Ca²⁺ and Mg²⁺) containing 10 mM HEPES. The testcompound or DMSO (control) was added again at the same concentrationsthat were used during cell loading. The cell suspension was aliquotedinto a 96-well reading plate (Corning) in a volume of 90 μL (10⁵cells/well). The compound plate contained agonist (CXCL8 in HBSS⁻) orHBSS⁻ (control). After 15 sec of reading the basal level of fluorescenceby FlexStation II, 10 μL of CXCL8 or HBSS⁻ were automaticallytransferred from the compound plate into the reading plate (finalconcentration of CXCL8 was 25 nM). Changes in fluorescence weremonitored (ex=485 nm, em=525 nm) every 5 s for 240 to 500 s at roomtemperature.

The maximum change in fluorescence, expressed in arbitrary units overbaseline (Max-Min), was used to determine the CXCL8 response. The effectof each compound on the CXCL8 response was normalized and expressed as apercent of the DMSO control, which was designated as “100% response.”Curve fitting and calculation of the compound inhibitory concentrationthat reduces the level of the CXCL8 response by 50% (IC₅₀), or thecompound agonist concentration that increases the level of the calciumrelease by 50% of the maximum agonist-induced change (EC₅₀) weredetermined by nonlinear regression analysis of the dose-response curvesgenerated using Prism 4 (GraphPad Software, Inc., San Diego, Calif.).

The mean (±SE) IC₅₀ for SX-682 (n=4) was 42±3 nM, 20±2 nM and 55±6 nM inCXCR1 transfected RBL cells (‘CXCR1’, squares), CXCR2 transfected RBLcells (‘CXCR2’, inverted triangles), and human neutrophils (‘HumanPMNs’, circles), respectively (FIG. 1).

Pharmacology Example 2: SX-682 Exhibits Sustained Wash-ResistantInhibition of Intracellular Calcium Release

SX-682 contains a boronic acid moiety that has the potential to form atransient covalent linkage with hydroxyl-bearing amino acid side chainsin the binding site of its protein target. Without wishing to be boundby theory, we hypothesized that such a transient covalent linkage in thebinding site of SX-682 might result in CXCR1/2 inhibition that wassustained after inhibitor washout. If inhibition is sustained in vitroafter SX-682 washout, inhibition may also be sustained in vivo afterSX-682 has been eliminated from the plasma, a property that would permitinfrequent patient dosing regimens (e.g. once-daily, twice-weekly andonce-weekly). Infrequent dosing regimens are preferred embodiments.

In order to test this hypothesis, RBL cells (10′ cells/mL) stablytransfected with either CXCR1 or CXCR2 were (1) incubated with SX-682 atvarious concentrations for 30 minutes at 37° C., (2) washed andresuspended in assay buffer (RPMI/2% serum) at room temperature, and (3)assayed for CXCL8-mediated calcium response at time points up to 12 hafter inhibitor washout. The concentrations of SX-682 tested were 0(positive control), 0 (negative control), 0.4, 2, and 10 μM. At 30minutes before each time point, a 56.25 μL aliquot of the cells wereremoved and loaded for 30 minutes at room temperature in the dark withFLIPR-3 reagent (262.5 μL per tube). Following FLIPR-3 incubation, cellswere assayed for CXCL8 mediated intracellular calcium release asdescribed in Pharmacology Example 1.

Consistent with our hypothesis, SX-628 exhibited inhibition ofCXCL8-mediated intracellular calcium flux in RBL cells stablytransfected with either CXCR1 (FIG. 2) or CXCR2 (FIG. 3) that wassustained for at least 12 hours after SX-682 washout.

Pharmacology Example 3: SX-682 Exhibits Pronounced Activity in the RatModel of Pulmonary Inflammation

SX-682 was assayed in an in vivo rat model of pulmonary inflammation.Activity in this model of pulmonary inflammation provides evidence thatsupports the use of SX-682 in the treatment of a number of pulmonaryinflammatory diseases, including chronic obstructive pulmonary disease(COPD) and bronchopulmonary dysplasia (BPD). In this experiment,Sprague-Dawley rats (n=4 per cohort) were dosed intravenously only onceat t=0 with either vehicle control (dimethylformamide/PEG400/saline,40:40:20), positive inhibitor control (SX-576, 1 mg/kg) or the testcompound (SX-682, 1 mg/kg). The rats were then placed in air (negativeexposure group; vehicle control only) or 1 ppm ozone (positive exposuregroup; vehicle control, positive inhibitor control SX-576, and testcompound SX-682) for 4 hours. The rats were then sacrificed at t=24hours, and the bronchoalveolar lavage fluid (BALF) was collected. Thecells were spun down, stained with Wright-Giemsa and counted. In thenegative exposure group, no neutrophils were observed when stained. Inthe ozone exposed rats treated with vehicle however, there was a briskinflux of neutrophils of approximately 14,000 per mL of BALF (FIG. 4).In contrast, both SX-576 and SX-682 (each at 1 mg/kg) significantlydecreased the influx of neutrophils into the lungs as compared tocontrol rats treated with vehicle only (FIG. 4). Of the two inhibitorstested, SX-682 exhibited a markedly more robust inhibition of neutrophilchemotaxis (FIG. 4). Notably, the inhibition of neutrophil influx intothe BALF was sustained for 24 hours after only a single dose of 1 mg/kgof SX-682. These results provide evidence that SX-682 is a potentinhibitor of pulmonary neutrophil chemotaxis in vivo, and is effectivefor treating diseases with a heightened pulmonary inflammationcomponent, like COPD in a predictive in vivo model.

Stability Example 1: Increased Microsomal Stability of SX-682

Liver microsomes are an in vitro model for in vivo metabolism andelimination of a drug by the liver (and gut) cytochrome P450 system. Acompound's stability in liver microsomes in vitro is predictive of itsmetabolism and elimination in vivo. The stability of SX-682 in livermicrosomes together with several other cogeners was examined to quantifythe microsomal stability of SX-682 and identify potentialstructure-activity relationships (SAR) predictive of stability orinstability (FIG. 5).

The compounds were incubated in duplicate with human liver microsomes at37° C. The reaction contained microsomal protein in 100 mM potassiumphosphate, 2 mM NADPH, 3 mM MgCl₂, pH 7.4. A control was run for eachcompound omitting NADPH to detect NADPH-independent degradation. Analiquot was removed from each experimental and control reaction at 0,10, 20, 30, and 60 minutes and mixed with an equal volume of ice-coldStop Solution (0.3% acetic acid in acetonitrile containing haloperidol,diclofenac, or other internal standard). Stopped reactions wereincubated for at least ten minutes at −20° C., and an additional volumeof water was added. The samples were centrifuged to remove precipitatedprotein, and the supernatants were analyzed by LCMS/MS to quantitate theremaining compound. Data were converted to percent remaining by dividingby the time zero concentration value. Data were fit to a first-orderdecay model to determine half-life. Intrinsic clearance was calculatedfrom the half-life and the protein concentrations:CL _(int)=ln(2)/(t½ [microsomal protein]).The results are shown in Table 1. Surprisingly, SX-682 was markedly morestable than SX-671 or SX-576 (6-fold larger half-life), even though thelatter is structurally identical but for a single ring nitrogen. On theother hand, the introduction of a ring nitrogen was insufficient aloneto impart the stability seen with SX-682 as demonstrated by SX-677 andSX-678, which have half-lives that are 2-fold and 5-fold smaller thanSX-682, respectively. More surprising is that eliminating the ringnitrogen in SX-517 yielded a half-life even larger than that of SX-682.The results as a whole led to no SAR predictive of the surprisingstability of SX-682.

TABLE 1 Stability in human liver microsomes (NADPH-dependent) ParameterSX-517 SX576 SX-671 SX-677 SX-678 SX-682 CL_(int) ^(a) (μL/ 3.4 45.716.2 15.9 33.4 2.1 min mg) t½^(b) (min) 405 50 143 145 69.2 325^(a)microsomal intrinsic clearance ^(b)half-life

Metabolic Stability Example 2: Increased Plasma Stability of SX-682

The in vitro stability of SX-682 and the cogeners of Metabolic StabilityExample 1 (FIG. 5) were further studied in human plasma. The reactionswere initiated by the addition of 5 μL of a 500 μM DMSO stock solutionto 495 μL of preheated plasma solution to yield a final concentration of5 μM. The assays were performed in a heat block at 37° C. and conductedin duplicate. Samples (50 μL) were taken at 0, 30, 60, 120, 240 min andadded to 150 μL acetonitrile in order to deproteinize the plasma. Thesamples were subjected to vortex mixing for 1 min and thencentrifugation for 15 min at 14,000 rpm. The clear supernatants wereanalyzed by LC-MS.

The in vitro plasma half-life (t½) was calculated using the expressiont½=ln(2)/bwhere b is the slope found in the linear fit of the natural logarithm ofthe fraction remaining of the parent compound vs. incubation time.

The results are shown in Table 2. In the case of plasma stability,SX-682 is roughly as stable as SX-576 in contradistinction to itsmarkedly enhanced stability in liver microsomes. Apparently, eliminatingthe ring nitrogen has little impact on plasma stability. On the otherhand, also changing the sulfur to oxygen in SX-671 resulted in apronounced 35-fold reduction in plasma half-life. However, keeping thesulfur is insufficient alone to maintain plasma stability as illustratedby SX-517, which maintains the sulfur but eliminates the ring F3CO groupand results in a 5-fold reduction in plasma half-life.

TABLE 2 Stability in human plasma (incubation at 37° C., LC-MS/MSdetection) Parameter SX-517 SX-576 SX-671 SX-677 SX-678 SX-682 t½ (min)113 533 21 2310 3465 693

Formulation Example 1: Spray Dry Dispersion (SDD) of SX-682 ontoHypromellose Phthalate (HPMCP)

An alternative method to increase oral bioavailability of drug productsis through spray dry dispersion in a polymer carrier matrix, whichutilizes the spray drying of the active pharmaceutical ingredient (API)and polymer in an organic solvent. Upon drying, the API is amorphouslydispersed in the polymer matrix. The polymer matrix is water soluble andallows for the slow release of API upon exposure to aqueousenvironments. Spray dry dispersion was performed at Emerson Resources(Norristown, Pa.). SX-682 (250 g) was first dissolved in acetone/water(97:3, 10 liters). HPMCP, 750 g, was then added. The in-process controlis to check the clarity of the feed solution and report the result. Thematerial is then spray dried using a GEA Mobile Minor spray dryer usingnitrogen as the drying gas. The inlet temperature is set at 90° C., andthe outlet temperature is 55-60° C. The nozzle size used was 1 mm. TheSX-682 spray solution concentration is 10%, and the atomization nitrogenwas set at 1.8 bar, 50%. The formulated powder is collected from boththe cyclone and the cartridge filter and post-dried. The powder isbag-blended and double bagged with dessicant pouches for storage. Thespray-dried dispersion was suspended in aqueous 0.5% methylcellulose(400 cps) prior to oral dosing in preclinical models.

Pharmacology Example 4: Inhibition of MDSC Migration

A large body of evidence indicates that chronic inflammation as occursin inflammatory bowel disease is one of several key risk factors forcancer initiation, progression, and metastasis. In anazoxymethane/dextran sulfate sodium (AOM/DSS) model ofcolitis-associated cancer, Katoh, 2013, Cancer Cell, 24:631-44,presented genetic evidence (Cxcr2^(−/−)) that loss of CXCR2 dramaticallysuppressed chronic colonic inflammation and colitis-associatedtumorigenesis by inhibiting MDSC recruitment into colonic mucosa andtumors. CXCR2 ligands CXCL1, CXCL2 and CXCL5 were all elevated ininflamed colonic mucosa and tumors and induced MDSC chemotaxis. Adoptivetransfer of wild-type MDSCs into Cxcr2^(−/−) mice restoredAOM/DSS-induced tumor progression. Deletion of Cxcr2 did not affectinfiltration of dendritic cells, T cells, NK or NKT cells. Deletion ofCxcr2 significantly decreased the migratory ability of MDSCs in vitroand in vivo. MDSCs accelerated tumor growth by inhibiting CD8+ T cellcytotoxic activity. Their results showed that CXCR2 was required forhoming of MDSCs into colonic mucosa and colitis-associated tumors,revealing a role of CXCR2 in the recruitment of MDSCs from thecirculatory system to local tissues and tumors. The results fromadoptive transfer of MDSCs provide direct evidence that MDSCs contributeto colonic tumor formation and growth by inhibiting CD8+ T cellcytotoxicity against tumor cells. The authors conclude, “our findingsprovide a rationale for the development of therapeutic approaches tosubvert . . . tumor-induced immunosuppression by using CXCR2 antagonists. . . ”

Antagonist SX-682 was used to validate the concept of using a smallmolecule antagonist of CXCR1 and CXCR2 receptors to block MDSCmigration. Using the of method of Katoh, 2013, Cancer Cell, 24:631-44(herein incorporated by reference), to induce and isolate MDSCs, maleBALBc mice were given a single intraperitoneal injection of azoxymethane(AOM, 10 mg/kg). Seven days later, these mice were given 4 cycles ofwater and water containing 1.25% dextran sulfate sodium (DSS). One cycleconsisted of providing mice with regular drinking water for 7 days,followed by 1.25% aqueous DSS for 7 days, followed by regular drinkingwater for 7 days. MDSCs were isolated from femurs and blood ofAOM/DSS-treated BALBc mice using the Myeloid-Derived Suppressor CellIsolation Kit (Miltenyi Biotec) after lysis of red blood cells (RBC)according to the manufacturer's instructions. GRO□□(CXCL1) was placed inthe bottom chamber of a 24-well plate at a concentration of 100 ng/ml.MDSCs isolated from blood of AOM/DSS-treated BALBc mice were seeded at adensity of 1×10⁶/well in the upper chamber (3 μm, BD Falcon). Afterincubation for 12 hours, migrated cells were quantified by flowcytometry using CountBright Absolute Counting Beads (Molecular Probes).To evaluate the effect of SX-682 on GRO□-mediated MDSC migration,aqueous DMSO solutions of SX-682 were added to the MDSC-seeded wellsprior to migration. The test concentrations of SX-682 were 0.1, 1 and 10□M, and the DMSO concentration was <1%. The results showed that SX-682was able to effectively inhibit GRO□-mediated MDSC migration in adose-dependent manner (FIG. 7).

Pharmacology Example 5: Inhibition of Tumor Cell Proliferation by SX-682in a Dose-Dependent Manner

The chemokine CXCL8 and its receptors CXCR1 and CXCR2 have beenidentified as important mediators of cellular proliferation for a numberof tumor cell types. To validate the relationship between CXCR1 andCXCR2 receptor antagonism and inhibition of tumor cell proliferation,SX-682 was evaluated against the 60 human tumor cell line panel at fiveconcentration levels (0.01, 0.1, 1, 10, 100 □M) as performed by theDevelopmental Therapeutics Program at the National Cancer Institute(Shoemaker, 2006, Nat Rev Cancer, 6:813-23).

The human tumor cell lines of the cancer screening panel were grown inRPMI 1640 medium containing 5% fetal bovine serum and 2 mM L-glutamine.Cells were inoculated into 96 well microtiter plates in 100 μL atplating densities ranging from 5,000 to 40,000 cells/well depending onthe doubling time of individual cell lines. After cell inoculation, themicrotiter plates were incubated at 37° C., 5% CO₂, 95% air and 100%relative humidity for 24 h prior to addition of SX-682.

After 24 h, two plates of each cell line are fixed in situ with TCA, torepresent a measurement of the cell population for each cell line at thetime of drug addition (Tz). SX-682 was solubilized in dimethyl sulfoxideat 400-fold the desired final maximum test concentration and storedfrozen prior to use. At the time of SX-682 addition, an aliquot offrozen concentrate was thawed and diluted to twice the desired finalmaximum test concentration with complete medium containing 50 μg/mlgentamicin. Additional four, 10-fold or ½ log serial dilutions are madeto provide a total of five drug concentrations plus control. Aliquots of100 μl of these different drug dilutions were added to the appropriatemicrotiter wells already containing 100 μl of medium, resulting in therequired final drug concentrations.

Following drug addition, the plates were incubated for an additional 48h at 37° C., 5% CO₂, 95% air, and 100% relative humidity. For adherentcells, the assay was terminated by the addition of cold TCA. Cells werefixed in situ by the gentle addition of 50 μl of cold 50% (w/v) TCA(final concentration, 10% TCA) and incubated for 60 minutes at 4° C. Thesupernatant was discarded, and the plates were washed five times withtap water and air dried. Sulforhodamine B (SRB) solution (100 μl) at0.4% (w/v) in 1% acetic acid was added to each well, and plates wereincubated for 10 minutes at room temperature. After staining, unbounddye was removed by washing five times with 1% acetic acid and the plateswere air dried. Bound stain was subsequently solubilized with 10 mMTRIS, and the absorbance was read on an automated plate reader at awavelength of 515 nm. For suspension cells, the methodology is the sameexcept that the assay was terminated by fixing settled cells at thebottom of the wells by gently adding 50 μl of 80% TCA (finalconcentration, 16% TCA). Using the seven absorbance measurements (timezero, (Tz), control growth, (C), and test growth in the presence of drugat the five concentration levels (Ti)), the percentage growth wascalculated at each of the drug concentrations levels. Percentage growthinhibition was calculated as:(Ti−Tz)/(C−Tz)×100 for concentrations for which Ti>/=Tz(Ti−Tz)/Tz×100 for concentrations for which Ti<Tz.

Three dose response parameters were calculated for each experimentalagent. Growth inhibition of 50% (GI50) is calculated from[(Ti−Tz)/(C−Tz)]×100=50, which was the drug concentration resulting in a50% reduction in the net protein increase (as measured by SRB staining)in control cells during the drug incubation. The drug concentrationresulting in total growth inhibition (TGI) is calculated from Ti=Tz. TheLC50 (concentration of drug resulting in a 50% reduction in the measuredprotein at the end of the drug treatment as compared to that at thebeginning) indicating a net loss of cells following treatment iscalculated from (Ti−Tz)/Tz×100=−50. Values are calculated for each ofthese three parameters if the level of activity is reached; however, ifthe effect is not reached or is exceeded, the value for that parameteris expressed as greater or less than the maximum or minimumconcentration tested.

The results (FIGS. 8A, 8B, 8C, 8D, 8E, 8F, 8G, 8H, and 8I) validated theability of SX-682 to directly affect the proliferation of human tumorcells in the absence of other cells (e.g., MDSCs) through CXCR1/2antagonism.

Pharmacology Example 6A: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Carboplatin in the T11 Mouse Model ofBreast Cancer

The T11 model is a validated mouse model of breast cancer, which derivesfrom the serial orthotopic transplantation of a murine breast tumorderived from a p53-null mouse into a syngeneic p53 competent recipient,and features sporadic, somatic K-Ras mutation (Herschkowitz, 2012, ProcNatl Acad Sci, 109:2778-83, herein incorporated by reference). Tumorsfrom the T11 model display an RNA expression pattern characteristic ofthe human claudin-low disease, and are extremely aggressive, with themajority of untreated animals surviving less than 21 days from the timeof enrollment in the therapy studies. Treatment regimens were startedfollowing tumor manifestation. SX-682 was given orally via medicatedfeed, at an approximate dose of 10 mg/day/mouse. In brief, the medicatedfeed was prepared by Research Diets (New Brunswick, N.J.) byincorporating 15.2 grams of SX-682 spray dried dispersion (as preparedin Formulation Example 1) into 1042 grams of standard rodent diet with10% kcals from fat. The medicated feed was formed into pellets.Carboplatin (CP) was administered once weekly via intraperitonealinjection at a dose of 50 mg/kg. The results (FIG. 9) showed theaddition of SX-682 to carboplatin significantly reduced tumor growth(N=7 each cohort). The combination of SX-682 and CP resulted in asynergistic inhibition of tumor growth and progression in treatmentanimals, which is a surprising and unexpected result. Mean±SE. ***P<0.0001 (linear regression).

Pharmacology Example 6B: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Carboplatin in the C3Tag Mouse Model ofBreast Cancer

The C31-T-antigen (C3Tag) mouse model is a validated model of humantriple-negative basal cell breast cancer as shown by gene expressionanalysis (Maroulakou, 1994, Proc Natl Acad Sci, 91:11236-40, hereinincorporated by reference). The expressed large T-antigen binds andinactivates the RB and p53 tumor suppressor genes, explaining why thismodel faithfully recapitulates human basal breast cancer, which alsoharbors RB and p53 inactivation. This model also has frequent K-Rasamplification and infrequent Ras mutations. Treatment regimens werestarted following tumor manifestation. SX-682 was given orally viamedicated feed, at an approximate dose of 10 mg/day/mouse. CP wasadministered once weekly via intraperitoneal injection at a dose of 50mg/kg. The results (FIGS. 10 and 11) showed SX-682 treatment alone andin combination with carboplatin significantly reduced tumor growth ascompared to untreated controls (N=10-12 per cohort). Mean±SE. ** P<0.001(linear regression). Median survival for vehicle, carboplatin and SX-682cohorts was 18, 28 and 28 days. Combining SX-682 with CP significantlyincreased survival compared to carboplatin alone (P=0.008), giving amedian survival well beyond 60 days. The combination of SX-682 and CPresulted in a synergistic extension of survival in treatment animals,which is a surprising and unexpected result.

Pharmacology Example 6C: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Carboplatin in the TRIA Mouse Model ofMelanoma

The Tyrosine-Hras/Ink/Arf null (TRIA) is a validated mouse model ofmelanoma features an immuno-competent autochthonous tumor; Ras eventsare in >70% of all melanomas, and ˜50% lose the INK4a/ARF locus which,with B-RAF mutations, is the most common lesion of this cancer(Sharpless, 2016, Cancer Cell, 29:832-45). Treatment regimens werestarted following tumor manifestation. SX-682 was given orally viamedicated feed, at an approximate dose of 10 mg/day/mouse. CP wasadministered once weekly via intraperitoneal injection at a dose of 50mg/kg. The results of this experiment can be seen in FIGS. 12 and 13.SX-682 treatment alone significantly slowed tumor growth comparable toCP alone. SX-682 combination therapy did better than monotherapy andachieved complete remission (zero tumor volume) in 5 of 11 animals. Thecombination of SX-682 and CP resulted in a synergistic inhibition oftumor growth and progression in treatment animals, which is a surprisingand unexpected result.

Pharmacology Example 6D: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Temozolomide in a Mouse Model ofGlioblastoma Multiforme

Mouse glioma cell line GL261 is frequently used in experimental modelsof glioblastoma to evaluate various experimental modalities. Szatmari,2006, Cancer Sci, 97:546-55 (herein incorporated by reference). To testthe hypothesis that dual CXCR1/2 antagonism may potentiate anti-tumoreffects of chemotherapy in a mouse model of glioblastoma, tumor-bearingC57BL/6 mice are dosed with CXCR1/2 antagonist alone and in combinationwith temozolomide (TMZ). Mouse syngeneic tumor GL261 cells are grownwith Dulbecco modified Eagle medium supplemented with 10% fetal bovineserum as well as streptomycin (100 mg/mL) and penicillin (100 U/mL) at37° C. in a humidified atmosphere of 95% air/5% CO₂. Mouse glioma GL261cells are cultured, harvested, and injected into the lower right flankof each C57BL/6 mouse. For the subcutaneous model, 10⁶ GL261 cells areinjected into the lower right flank of each C57BL/6 mouse. TMZ is dosedvia intraperitoneal injection according to body weight (20 mg/kg).CXCR1/2 antagonist is dosed via oral administration daily. At thebeginning of treatment, mice are either randomized by tumor volume or bybody weight. The number of animals per group range from between 10-12animals per group as determined based on Good Statistical Practiceanalysis. Both tumor and body weight measurements are collected twiceweekly and tumor volume is calculated using the equation (L×W²)/2, whereL and W refer to the length and width dimensions, respectively. Errorbars are calculated as standard error of the mean. The general health ofmice is monitored daily and all experiments are conducted in accordanceto AAALAC and institution-based IACUC guidelines for humane treatmentand care of laboratory animals. Kaplan-Meier statistical analysis isperformed using the Log-rank test using GraphPad Prism.

Pharmacology Example 7A: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Anti-PD1 Antibody in the B16-F10Syngeneic Mouse Model of Melanoma

The B16-F10 mouse model is an established model of melanoma, Overwijk,2001, CURR PROTOC IMMUNOL, Chapter 20:Units 20-21 (herein incorporatedby reference), which was used to evaluate the effect of SX-682 alone andin combination with anti-PD1 antibody. B16-F10 mouse melanoma cells werecultured, then mice were injected with 0.5×106 B16-F10 cells on day 0,and treatment was initiated on day 18. Mice were treated with vehicle(control), SX-682 alone (50 mg/kg twice daily, oral), anti-PD-1 alone(100 μg twice weekly, ip), or SX-682 in combination with anti-PD-1(N=4-5 per cohort). SX-682 was used as a spray dried dispersion(Formulation Example 1) and was administered to mice via oral gavage asa suspension in 0.5% aq methylcellulose. The results (FIG. 14) showedSX-682 monotherapy significantly slowed tumor growth (P=0.0002, linearregression) and potently synergized with anti-PD1 therapy with thecombination significantly better than either therapy alone (P<0.0005 forboth comparisons). Data and error bars are the mean±SE. The combinationof SX-682 and immune checkpoint inhibition resulted in a synergisticinhibition of tumor growth and progression in treatment animals, whichis a surprising and unexpected result.

Pharmacology Example 7B: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Anti-PD1 Antibody in the LLC SyngeneicMouse Model of Lung Cancer

The Lewis lung carcinoma (LLC) mouse model (Kellar, 2015, Biomed ResInt, 2015:621324, herein incorporated by reference) was used to evaluatethe effect of SX-682 alone and in combination with anti-PD-1 antibody.LLC mouse melanoma cells were cultured, then mice were injected with0.5×106 LLC cells on day 0, and on day 18 treatment was initiated. Micewere treated with vehicle (control), SX-682 alone (50 mg/kg twice daily,oral), anti-PD-1 alone (100 μg twice weekly, ip), or SX-682 incombination with anti-PD-1 (N=5 per cohort). SX-682 was used as a spraydried dispersion (Formulation Example 1) and was administered to micevia oral gavage as a suspension in 0.5% aq. methylcellulose. The resultsof this experiment can be seen in FIGS. 15 and 16. SX-682 monotherapysignificantly slowed tumor growth (P=0.0076, linear regression), andpotently synergized with anti-PD1 therapy with the combinationsignificantly better than either therapy alone (P<0.0001 for bothcomparisons). The combination of SX-682 and anti-PD-1 therapysignificantly enhanced survival compared to vehicle or anti-PD-1 therapyalone (P=0.002). The combination of SX-682 and immune checkpointinhibition resulted in a synergistic effect on both tumor growth andsurvival in treatment animals, which is a surprising and unexpectedresult.

Pharmacology Example 7C: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with Anti-PD1 and Anti-CTLA4 Antibody in aMouse Model of Castration Resistance Prostate Cancer

The genetically engineered Ptenpc−/−p53pc−/−Smad4pc−/− mouse is aprostate-specific PB promoter-driven (PB-Cre4) conditional tripleknockout model (i.e., prostate-specific deletion of all three tumorsuppressors, Pten, p53 and Smad4 occurs in the ‘prostate cancer’ or‘pc’). It exhibits an aggressive tumor phenotype, and like the humandisease, develops spontaneous bone metastases (Ding, 2012, Cell,148:896-907) Ptenpc−/− p53pc−/−Smad4pc−/− mice at 3-4 months of age withestablished tumors (>150 mm3, as measured by MRI) were treated witheither vehicle (control), SX-682 (50 mg/kg b.i.d.), ICB (200 μg each ofanti-PD1 and anti-CTLA4 antibodies, 3×/week), or the SX-682 and ICBcombination (SX-682+ICB). SX-682 was used as a spray dried dispersion(Formulation Example 1) and was administered to mice via oral gavage asa suspension in 0.5% aq. methylcellulose. Treatment was continued for4-6 weeks and prostate weights measured to determine tumor burden. Theresults (FIG. 17) showed SX-682 plus ICB was significantly better thancontrol or either treatment alone (unpaired t-test). Mean+SE. Thecombination of SX-682 and immune checkpoint inhibition resulted in asynergistic inhibition of tumor growth and progression in treatmentanimals, which is a surprising and unexpected result.

Pharmacology Example 8A: Cell-Kill Efficacy of CXCR1/2 Antagonism asSingle Therapy and in Combination with T-Cell Therapy In Vitro

To validate the use of a small molecule CXCR1/2 antagonist to halt theEMT process in tumor cells thereby decreasing immunoresistance, humantumor cells will be exposed in vitro to various doses of SX-682 in thepresence of immune effector cells. These studies will be conducted withand without addition of exogenous recombinant IL-8. Tumor cells will besubsequently evaluated for proliferation/survival, expression ofepithelial/mesenchymal markers and stemness markers, expression ofimmune-relevant molecules, and cytotoxic response to various immuneeffector cells (human antigen-specific T cell lines generated from theblood of cancer patients or healthy donors by using peptide epitopesfrom the following antigens: CEA, MUC1, brachyury, or natural killercells isolated from blood of normal donors and left untreated oractivated via IL-2)

Pharmacology Example 8B: Efficacy of CXCR1/2 Antagonism as SingleTherapy and in Combination with T-Cell Therapy in a Mouse Model ofPancreatic Cancer

Although born with histologically normal pancreata, K-rasLSL.G12D/+;p53R172H/+; PdxCre (KPC) mice developed pancreatic intraepithelialneoplasia (PanIN) lesions on an accelerated schedule and died ofpancreatic ductal adenocarcinoma (PDAC) with a median survival of 5.5months. Metastases were observed ˜80% of the animals, at the same sitesseen in human PDAC patients (liver, lung, and peritoneum). Tumorsarising in this model were found to have many of the immunohistochemicalmarkers associated with human disease, and bore evidence of widespreadgenomic alterations, a feature that was previously lacking in mostgenetically engineered mouse models. The KPC mouse model of PDAC iswidely used to evaluate treatment modalities in a preclinical setting(Westphalen, 2012, Cancer J, 18:502-10, incorporated herein byreference). To test the hypothesis that dual CXCR1/2 antagonism maypotentiate anti-tumor effects of T-cell therapy in a mouse model ofPDAC, tumor-bearing KPC mice are dosed with SX-682 alone and incombination with T-cell therapy. T-cells are either selected for orengineered towards high affinity binding to a specific protein ofinterest located on the tumor cell. In the case of PDAC, these proteinsof interest may include (but are not limited to) mesothelin, Wilms'tumor antigen, Mucin 1, or Annexin A2. KPC mice will undergo serialhigh-resolution ultrasound imaging (Vevo 2100) at 8 weeks of age tomonitor autochthonous tumor development. Mice are enrolled based ondefined pancreatic mass 3-6 mm. At the start of treatment, selectcohorts will receive CXCR1/2 antagonist via oral administration daily.For animals undergoing T-cell therapy, they will receivecyclophosphamide once at enrollment (180 mg/kg) followed 6 hours laterby intravenous infusion of 1×107 twice-stimulated engineered T cellsfollowed by 1×107 peptide-pulsed irradiated splenocytes. Recipient micewill also receive IL-2 (2×104 IU, i.p.) every other day for 8 days (intimepoint studies) or for 5 days (in survival studies) to promote donorT cell survival and expansion. For serial T cell infusions, mice willreceive the same treatment protocol, excluding the cyclophosphamideafter the first dose, every 2 weeks. Power analyses will guideenrollment numbers to power the study for a large effect (>50% increasein median overall survival). Both tumor and body weight measurements arecollected twice weekly and tumor volume is measured via serialhigh-resolution ultrasound imaging. Error bars are calculated asstandard error of the mean. The general health of mice is monitoreddaily and all experiments are conducted in accordance to AAALAC andinstitution-based IACUC guidelines for humane treatment and care oflaboratory animals. Kaplan-Meier statistical analysis is performed usingthe Log-rank test using GraphPad Prism.

Pharmacology Example 9: Efficacy of CXCR1/2 Antagonism as Single Therapyand in Combination with Cancer Vaccine in a Mouse Model of Cancer

All animal studies re carried out in accordance with the guidelines ofthe Association for Assessment and Accreditation of Laboratory AnimalCare. Experimental studies were carried out under approval of the NIHIntramural Animal Care and Use Committee. Murine colon carcinoma MC38cells are stably transfected with chorioembryonic antigen (CEA), andsubcutaneously implanted into female C57BL/6 mice on day 0. Beginning onday 7, animals are dosed daily with SX-682 via medicated feed. Beginningon day 14, test animals are vaccinated weekly with either Hank'sBalanced Salt Solution or 50 mg of a gp70 peptide (p15e) emulsified inMontanide ISA-51-VG (Seppic) at a 1:1 ratio. Evaluation of the effect ofSX-682 on various immune cell subsets in non-tumor bearing mice will beevaluated. Spleens will be collected and analyzed for antigen-specificimmune responses and various immune cell subsets. Anti-tumor effect ofcombinations of SX-682 with cancer vaccines will be evaluated.Evaluations will include effect on tumor volume, tumor microenvironment,including tumor phenotype, etc., will be evaluated. Vaccine-specificimmune responses will also be measured.

We claim:
 1. A method for treating a patient with a cancer, comprisingassaying levels of (a) CXCR1 and/or CXCR2 ligands or (b) myeloid-derivedsuppressor cells (MDSCs) and/or neutrophils in a patient with a cancer,and if the patient has increased levels of (a) or (b), wherein theincreased levels are caused by the cancer, then administering apharmaceutical composition comprising a therapeutically effective amountof a compound having a structure selected from the group consisting offormulas SX-517, SX-576, and SX-682,

or a pharmaceutically acceptable salt or solvate thereof.
 2. The methodof claim 1, further comprising administering an anticancer therapy. 3.The method of claim 2, wherein the anticancer therapy is a chemotherapy.4. The method of claim 2, wherein carboplatin is administered to thepatient.
 5. The method of claim 2 wherein said anticancer therapy isradiation therapy.
 6. The method of claim 1 wherein the compound has thestructure of formula SX-682 and the cancer is selected from cancers ofthe prostate, colorectum, pancreas, cervix, stomach, endometrium, brain,liver, bladder, ovary, testis, head, neck, skin, mesothelial lining,blood, esophagus, breast, muscle, connective tissue, lung, adrenalgland, thyroid, kidney, lymphoid system, bone marrow or bone,glioblastoma, mesothelioma, renal cell carcinoma, gastric carcinoma,sarcoma, choriocarcinoma, cutaneous basocellular carcinoma, andtesticular seminoma.
 7. The method of claim 1 wherein the compound hasthe structure of formula SX-682 and the cancer is selected from cancersof the prostate, pancreas, stomach, brain, liver, head, neck, skin,blood, breast, lung, glioblastoma, and mesothelioma.
 8. The method ofclaim 1 wherein the compound has the structure of formula SX-682 and thecancer is breast cancer.
 9. The method of claim 8, wherein thepharmaceutical composition is administered orally.
 10. The method ofclaim 9, further comprising administering a platinum chemotherapy. 11.The method of claim 1 wherein the compound has the structure of formulaSX-682 and the cancer is a melanoma.
 12. The method of claim 11, whereinthe pharmaceutical composition is administered orally.
 13. The method ofclaim 12, further comprising administering a platinum chemotherapy. 14.The method of claim 1, wherein the cancer is a lung cancer.
 15. Themethod of claim 14, wherein the pharmaceutical composition isadministered orally.
 16. The method of claim 1 wherein the compound hasthe structure of formula SX-682 and the cancer is a prostate cancer. 17.The method of claim 16, wherein the pharmaceutical composition isadministered orally.
 18. The method of claim 1, wherein the cancer is aglioblastoma.
 19. The method of claim 18, wherein the pharmaceuticalcomposition is administered orally.
 20. The method of claim 19, furthercomprising administering temozolomide.
 21. The method of claim 1,wherein the cancer is a pancreatic cancer.
 22. The method of claim 21,wherein the pharmaceutical composition is administered orally.
 23. Amethod for treating a cancer, comprising administering to a patient withthe cancer and with an increased level of (a) CXCR1 and/or CXCR2 ligandsor (b) myeloid-derived suppressor cells (MDSCs) and/or neutrophils,wherein the increased levels are caused by the cancer, a pharmaceuticalcomposition comprising a therapeutically effective amount of a dualCXCR1/2 antagonist selected from the group consisting of formulasSX-517, SX-576, and SX-682,

or a pharmaceutically acceptable salt or solvate thereof.
 24. The methodof claim 23, further comprising administering an anticancer therapy. 25.The method of claim 24, wherein the anticancer therapy is achemotherapy.
 26. The method of claim 25, wherein the chemotherapyincludes the step of administering carboplatin to the patient.
 27. Themethod of claim 24, wherein the anticancer therapy is radiation therapy.28. The method of claim 23, wherein the dual CXCR1/2 antagonist isnavarixin and the cancer is selected from cancers of the prostate,colorectum, pancreas, cervix, stomach, endometrium, brain, liver,bladder, ovary, testis, head, neck, skin, mesothelial lining, blood,esophagus, breast, muscle, connective tissue, lung, adrenal gland,thyroid, kidney, lymphoid system, bone marrow or bone, glioblastoma,mesothelioma, renal cell carcinoma, gastric carcinoma, sarcoma,choriocarcinoma, cutaneous basocellular carcinoma, and testicularseminoma.
 29. The method of claim 28, further comprising administering aplatinum chemotherapy.
 30. The method of claim 29, further comprisingadministering temozolomide.
 31. The method of claim 23, wherein thepharmaceutical composition is administered orally.